I was wondering if anybody knew any reason why primary cells when isolated did not have the ability to grow on regular polystyrene culture vessles- Sarstedt-red cap, and thus were grown on collagen coated vessles -sartedt-yellow cap.
However separate populations of these primary cells were immortalised (using one of two plasmids pSV3-neo and pRITA) and when subcultured back into red cap flasks they had no problem attaching.
My limited knowledge hypothesised that the primary cells lacked sufficent amounts of extra cellular matrix proteins to attach and this is why the attached and grew on the collagen coated flasks, however this does not explain why they would attach to the non-coated flasks when immortalised, or does it?
I mean i dont think the immortalisation step would have any bearing on extracellular matrix proteins or attchment rates so why would they attach to the red cap vessels now?
Also i should add the primary cells when isolated were over 200-250 um in diameter and when immortalised the their size reduced to 50-100 um.
Any suggestions or possible explanations would be great.
Edited by cotchy, 02 March 2010 - 08:27 AM.